Fenugreek 4 - Hydroxy-isoleucine Extraction And Separation Of Seed Utilization,

This paper studied the extraction, separation and purification of 4-hydroxyisoleucine and examination of the 4-hydroxyisoleucine content. and tentative research comprehensive utilization of fenugreek seed. The main results were as following:1. By orthogonal experiments, it is confirmed that the optimal extraction conditions :Hydrothermal refluxing extraction with the 90% ethanol ratio at 1/5,80℃lasting for four hours.2. To research the effects on 4-hydroxyisoleucine recovered from Trigonella foenum-graecum by ion exchange. Concentrated liquid PH=3, flow-rate 1BV/h, 0.5mol/L NH₃H₂O as eluant. The total recovery ratio is 62.63%, the purity is 45.36%.3. Purification technics of 4-hydroxyisoleucine:After silica gel column chromatography, the purity is 56. 21%, the recovery rate is 90.83%;After dry up silica gel column chromatography, the purity is 79.36%, the recovery is 63.91%;After lactonization, the purity is 84. 91%, the recovery is 98.33%.4. To determine 4-hydroxyisoleucine by TLC.developing solvent : chloroform-methanol-water(10:5:1), coloration:2mg/ml ninhydrin ethanol solution, the detection wavelength was 570nm, Y=2E+07x+26009(r=0.9981) ,the average recovery was 100. 3%, RSD=0.93%.5. The above used ion-exchange colophony was basified with 10% NH₃H₂O. After chloroform eluted, we gained alkaloids 0.0586g;After hydrothermal refluxing extraction, we gained alkaloids 0. 0476g.6. The result of the extracted fenugreek gum from residue: After hydrothermal extraction, the purity is 32. 24%, the yield is 7. 77%;After water extraction, the purity is 46.50%, the yield is 10.16%, After deproteinated with Sevage method, the purity is 58. 73% the yield is 22. 08%; After deproteinated with the improved Sevage method, the purity is 66.91% the yield is 22.39%; After deproteinated with chloroactic acid method, the purity is 54.62% the yield is 38.23%.7. The result of comprehensive utilization:Mature seeds were ground and defatted, we got fenugreed oid. The powder was extracted with 90% ethanol, after vacuum concentration, the total free amino acids were fixed on an ion-exchange colophony and eluted with NH₃H₂O, the ammonia solution was concentrated and lyophilized, we got 4-hydroxyisoleucine. Unabsorptive liquid is flavone and steroidal saponins, the used ion-exchange colophony was marinated with ammonia and then extracted with chloroform, we got alkaloids. Residue was extracted with water and then concentrated, we got fenugreek gum.